KNJIGA POVZETKOV BOOK OF ABSTRACTSvabljeno predavanje (IL) kratko predavanje (SL) poster (P) Predavatelji bodo imeli za svojo predstavitev osebni računalnik (Windows 7 in Office 2013)

6. SLOVENSKI SIMPOZIJ O RASTLINSKI BIOLOGIJI z mednarodno udeležbo Fakulteta za kmetijstvo in biosistemske vede 11. - 12. september 2014, Hoče

6. SLOVENSKI SIMPOZIJ O RASTLINSKI BIOLOGIJI z mednarodno udeležbo Fakulteta za kmetijstvo in biosistemske vede 11. - 12. september 2014, Hoče

6th SLOVENIAN SYMPOSIUM ON PLANT BIOLOGY with international participation

Faculty of Agriculture and Life Science

September 11 - 12, 2014, Hoče, Slovenia

KNJIGA POVZETKOV

BOOK OF ABSTRACTS

Uredniki / Edited by: Jasna Dolenc Koce Tine Grebenc Andreja Urbanek Krajnc Izdalo / Issued by: Slovensko društvo za biologijo rastlin Slovenian Society of Plant Biology Oblikovanje / Designed by: Jasna Dolenc Koce

CIP - Kataložni zapis o publikaciji Narodna in univerzitetna knjižnica, Ljubljana 581(082)(0.034.2) SLOVENSKI simpozij o rastlinski biologiji z mednarodno udeležbo (6 ; 2014 ; Hoče) Knjiga povzetkov [Elektronski vir] = Book of abstracts / 6. slovenski simpozij o rastlinski biologiji z mednarodno udeležbo,11.-12. september 2014, Hoče = 6th Slovenian Symposium on Plant Biology with International Participation, September 11-12, 2014, Hoče, Slovenia] ; [organizator] Slovensko društvo za biologijo rastlin = [organized by] Slovenian Society of Plant Biology ; [uredniki Jasna Dolenc Koce, Tine Grebenc, Andreja Urbanek Krajnc]. - Ljubljana : Slovensko društvo za biologijo rastlin = Slovenian Society of Plant Biology, 2014 ISBN 978-961-91014-4-5 1. Dolenc Koce, Jasna 2. Slovensko društvo za biologijo rastlin 275284224

KNJIGA POVZETKOV

I

ORGANIZACIJSKI ODBOR

Tine GREBENC, predsednik / President

Andreja URBANEK-KRAJNC, tajnica / Secretary

Aleš KLADNIK, blagajnik / Treasurer

Jasna DOLENC KOCE, program / Programme

Tina DEMŠAR, stik s pokrovitelji / Contact wit sponsors

Matevž LIKAR, spletna stran / Web page

NASLOV ORGANIZACIJSKEGA ODBORA

Organizacijski odbor 6. slovenskega simpozija o rastlinski biologiji / Organizing committee of the 6th Slovenian Symposium on Plant Biology Slovenian Forestry Institute (dr. Tine Grebenc) Večna pot 2 SI-1000 LJUBLJANA E-mail: [email protected] http://www.plantslo.org

mailto:[email protected]://www.plantslo.org/

II

POKROVITELJI

Zahvaljujemo se naslednjim pokroviteljem, ki so s svojo finančno pomočjo omogočili izvedbo simpozija.

Mikro+Polo d.o.o.

Fakulteta za kmetijstvo in biosistemske vede Univerze v Mariboru

Nacionalni inštitut za biologijo

Biotehniška fakulteta Univerze v Ljubljani

Kemomed d.o.o.

Mediline d.o.o.

Mettler Toledo d.o.o.

Omega d.o.o.

KRKA, tovarna zdravil, d.d.

Lotrič d.o.o.

CHEMASS d.o.o.

Kambič Laboratorijska oprema d.o.o.

III

SPLOŠNE INFORMACIJE Simpozij se bo začel v četrtek, 11. septembra 2014 ob 9.00 uri in končal v petek, 12. septembra 2014 popoldan. Simpozij bo potekal na Fakulteti za kmetijstvo in biosistemske vede, Pivola 10, 2311 Hoče pri Mariboru (Lat.:46.505466; Lon.:15.622579) Recepcija bo odprta vsak dan od 8.00 do 12.00 ure. Oblike prispevkov:

vabljeno predavanje (IL)

kratko predavanje (SL)

poster (P) Predavatelji bodo imeli za svojo predstavitev osebni računalnik (Windows 7 in Office 2013) in ozvočenje (1x fiksni mikrofon, 3x prenosni mikrofon). Predavatelje prosimo, da svoj material oddajo operaterju 15 minut pred začetkom prve sekcije dneva. Posterji bodo razstavljeni v osrednjem hodniku Fakultete za kmetijstvo in biosistemske vede (1. nadstropje novogradnje). Avtorje prosimo, da posterje obesijo na začetku simpozija in odstranijo po zaključku, da bodo na ogled celoten čas simpozija. Prosimo, da je v času, ki je v programu predviden za predstavitev, ob posterju navzoč vsaj eden od avtorjev. Udeleženci simpozija imajo možnost objave znanstvenega članka po opravljeni recenziji v naslednjih revijah:

Acta biologica Slovenica (http://bijh-s.zrc-sazu.si/ABS/EN/ABS/Index.asp)

Acta agriculturae Slovenica (http://aas.bf.uni-lj.si/index-en.htm)

Acta silvae et Ligni (http://www.forestry.bf.uni-lj.si/) Kotizacija vključuje udeležbo na predavanjih, knjigo povzetkov, osvežitve med odmori, kosilo in slavnostno večerjo.

IV

The symposium will start on Thursday, September 11th 2014 at 9.00 a.m. and will close on Friday, September 12th 2014 in the afternoon. The Symposium will be held at the Faculty of Agriculture and Life Sciences, Pivola 10, SI-2311 Hoče near Maribor, Slovenia (Lat.:46.505466; Lon.:15.622579). The Symposium reception desk will be open every day from 8.00 to 12.00. Contributions will be presented as:

invited lecture (IL)

short lecture (SL)

poster (P) Facilities for projecting slides (personal computer equipped with Windows 7 in Office 2013) and audio system will be available. The speakers are kindly requested to deliver their presentation material at least 15 minutes prior the beginning of the first session of the day at the operator's desk. Posters will be displayed in the main corridor (1st floor) of the Faculty of Agriculture and Life Sciences. Authors are kindly requested to mount their posters at the beginning of the Symposium and to dismount them after the last session in order to present their work throughout the period of the Symposium. At least one of the authors is requested to be available for discussion during the corresponding poster session (indicated in the program). Scientific papers can be submitted for the publication in following peer reviewed journals:

Acta biologica Slovenica (http://bijh-s.zrc-sazu.si/ABS/EN/ABS/Index.asp)

Acta agriculturae Slovenica (http://aas.bf.uni-lj.si/index-en.htm)

Acta silvae et Ligni (http://www.forestry.bf.uni-lj.si/) Registration fee includes admission to all scientific sessions, book of abstracts, refreshments during breaks, lunch and conference dinner.

V

URNIK 11.9.2014 12.9.2014

Sept 11, 2014 Sept 12, 2014

Otvoritev

9:00 Opening

Moderne tehnologije

Modern technologies

9:30

Sistemska in molekularna

Systems and molecular biologija rastlin

10:00 plant biology

Rastlinska biotehnologija 10:30

Plant biotechnology

odmor

break

odmor 11:00

break

11:30

Sistemska in molekularna Struktura in funkcija

Systems and molecular Structure and function

biologija rastlin rastlin 12:00 plant biology

postri 12:30

poster session

kosilo 13:00

lunch

kosilo

lunch

13:30

14:00

postri

poster session

Interakcije rastlin 14:30

Plant interactions

z drugimi organizmi

with other organisms

15:00

Plant interactions

Interakcije rastlin z okoljem

15:30 with environment

odmor

break

16:00

odmor Interakcije rastlin

break Plant interactions

z drugimi organizmi 16:30

with other organisms

17:00

Interakcije rastlin z okoljem Dodiplomska sekcija

Plant interactions Undergraduate session

17:30 with environment

Zaključek 18:00

Closing

Večerja

18:30 Conference dinner

VI

PROGRAM SIMPOZIJA

Četrtek, 11. 9. 2014 / Thursday, September 11th 2014

08.00 – 09.00 Registracija / Registration

09.00 - 09.30 Otvoritev / Opening ceremony

1 Sistemska in molekularna biologija rastlin

Predsedujoči / Chair: P. Tijskens, K. Gruden

09.30 - 10.00 M. Naseem: A systems biology perspective on cytokinins crosstalk to plant immune networks (IL-1)

10.00 - 10.30 S. Pollmann: Induced auxin overexpression reveals a link between auxin signaling and lignification through the induction of ethylene biosynthesis and several cell wall modifying enzymes (IL-2)

10.30 - 10.45 N. Gozukirmizi: Sukkula retrotransposon insertion polymorphisms in barley calli (SL-1)

10.45 - 11.15 Odmor / Break

11.15 - 11.45 S. F. Elena: A systems biology approach to the evolution of virus-plant interactions (IL-3)

11.45 - 12.15 Š. Baebler: Digging potato molecular responses to virus infection (IL-4)

12.15 - 12.30 D. Dobnik: Suitability of wild potato relatives for functional analysis of potato genes using virus induced gene silencing (SL-2)

12.30 - 12.45 A. Coll Rius: Functional analysis of an Ethylene Response Factor involved in potato defence response to PVY (SL-3)

12.45 - 13.00 A. Lazar: Involvement of potato (Solanum tuberosum L.) MKK6 in response to Potato virus Y (SL-4)

13.00 – 13.15 S. Suomalainen: Using a magnetic particle system in plant NA purification (presentation of ThermoFisher Scientific)

VII

13.15 - 14.15 Kosilo / Lunch 14.15 - 15.00 Poster sekcija / Poster session

(vse sekcije / all sessions)

2 A Interakcije rastlin z okoljem in drugimi organizmi

Predsedujoči / Chair: H. Pfanz, D. Vodnik

15.00 - 15.30 M. Müller: Dynamic compartment specific changes in glutathione and ascorbate levels in plants under different environmental conditions (IL-1)

15.30 - 15.45 S. Al Sayegh Petkovšek: Risk assessment of receptor organisms: analysis of intake of metals through food chain (SL-1)

15.45 - 16.00 Z. Jeran: Biomonitoring of temporal and spatial patterns of metal deposition in Slovenia during the last twenty years (SL-2)

16.15 - 16.30 I. Maček: Soil abiotic factors determine arbuscular mycorrhizal fungal communities in hypoxic soils at natural CO2 springs (mofettes) (SL-3)

16.15 - 16.45 Odmor / Break

16.45 - 17.15 J. Šuštar-Vozlič: Abiotic stress in agricultural plants: Phaseolus sp. and drought (IL-2)

17.15 - 17.30 R. Povše: Physiological studies of drought stress in hop (Humulus lupulus L.) (SL-4)

17.30 - 17.45 A. Golob: UV radiation and selenium addition affects silicon accumulation in buckwheat plants (SL-5)

17.45 - 18.00 T. Trošt Sedej: Growth in conifer and deciduous trees under enhanced UV-B radiation (SL-6)

18.00 - 18.15 K. Eler: The functioning of two karst ecosystems revealed by eddy-covariance measurements (SL-7)

18.30 - Slavnostna večerja / Conference dinner

VIII

Petek, 12. 9. 2014 / Friday, September 12th 2014

3 Moderne tehnologije in Rastlinska biotehnologija

Predsedujoči / Chair: S. Pollman, J. Šuštar Vozlič

09.00 - 09.30 L. M. M. Tijskens: Green mathematics: Benefits of including biological variation in your data analysis (IL-1)

09.30 - 10.00 M. Kokalj: Infrared spectroscopy in plant science (IL-2)

10.00 – 10.30 D. Eichert: 3D Investigations of morphological and structural parameters in plants by Synchrotron Radiation X-ray micro Computed-Tomography (IL-3)

10.30 - 10.45 J. Ambrožič-Dolinšek: Cryopreservation by encapsulation-dehydration affects the morphology and regeneration capacity of endemic Hladnikia pastinacifolia (Apiaceae) (SL-1)

10.45 - 11.00 B. Bohanec: Report on the progress of development of biotechnological approaches for breeding pumpkins (Cucurbita pepo L.) (SL-2)

11.00 – 11.30 Odmor / Break

4 Struktura in funkcija rastlin

Predsedujoči / Chair: D. Eichert, A. Kladnik

11.30 – 12.00 H. Pfanz: Non-foliar photosynthesis - existence in and necessity for plants (IL-1)

12.00 – 12.15 M. Chersicola: Assymetry of molecular processes during abscission in tomato (Solanum lycopersicum L.) (SL-1)

12.15 – 12.30 T. Mrak: Arsenate and phosphate relationships during the uptake in lichens (SL-2)

12.30 - 13.00 Poster sekcija / Poster session

(vse sekcije / all sessions)

13.00 - 14.00 Kosilo / Lunch

IX

2 B Interakcije rastlin z okoljem in drugimi organizmi

Predsedujoči / Chair: M. Mueller, M. Naseem, J. Dolenc Koce

14.00 - 14.30 M. Likar: Distribution and diversity of arbuscular mycorrhizal fungi in grapevines from production vineyards along the eastern Adriatic coast (IL-1)

14.30 – 14.45 M. Flajšman: Testing of Verticillium albo-atrum virulence on different plant hosts and development of a transformation system for Verticillium albo-atrum gene knock-out (SL-1)

14.45 – 15.00 V. Progar: Transcriptional analysis of differential gene expression in hop following infection with Verticillium albo-atrum (SL-2)

15.00 – 15.15 T. Pokorn: Viroid-derived small RNA (vd-sRNA) target identification in hop (SL-3)

15.15 – 15.30 D. Kutnjak: What can "plant defence molecules" tell us about population structure of plant virus? (SL-4)

15.30 – 15.45 M. Pompe-Novak: Morphology and gene expression in lesions in compatible and incompatible potato – potato virus Y interaction (SL-5)

15.45 - 16.15 Odmor / Break

16.15 – 16.30 M. Petek: Potato Virus Y infection hinders potato defence response and renders plants more vulnerable to Colorado potato beetle attack (SL-6)

16.30 – 16.45 M. Rupar: Fluorescently labelled Potato virus Y (PVY): in vivo tracking and measuring the rate of spread of PVY in the host plants (SL-7)

16.45 – 17.00 J. Lamovšek: Pathogenic Agrobacterium tumefaciens antagonises root-knot nematode Meloidogyne ethiopica reproduction on tomato roots (SL-8)

17.00 - 18.00 Dodiplomska sekcija / Undergraduate session

Predsedujoči / Chair: M. Kokalj, S. Elena, T. Grebenc

Kratka predstavitev posterjev / Short presentation of posters Ocenjevanje / Evaluation

18.00 - 18.30 Zaključek / Closing

Razglasitev rezultatov Dodiplomske sekcije / Announcement of the results in the Undergraduate session

X

KAZALO

Sistemska in molekularna biologija rastlin 1 Plant systems biology and molecular plant biology Interakcije rastlin z okoljem in drugimi organizmi 14 Plant interactions with environment and other organisms Moderne tehnologije in Rastlinska biotehnologija 33 Modern technologies and Plant biotechnology Struktura in funkcija rastlin 45 Structure and function of plants Interakcije rastlin z okoljem in drugimi organizmi 53 Plant interactions with environment and other organisms Dodiplomska sekcija 70 Undergraduate session

Indeks avtorjev 82 Author indeks

Seznam udeležencev 86 List of participants

1

1 Sistemska in molekularna biologija rastlin

2

A systems biology perspective on cytokinins crosstalk to plant immune networks

Muhammad Naseem*, Thomas Dandekar Functional Genomics and Systems Biology group, Department of Bioinformatics, Biocenter, Am Hubland, D-97074 Wuerzburg, Germany * corresponding author ([email protected])

Molecular genetic studies have helped to elucidate a multistep two-component cytokinin (CK) signalling system in Arabidopsis. The importance of CK signalling in controlling plant growth and development has thoroughly been investigated. The efforts highlighting its implications in plant immunity are lagging behind. Higher plant CK levels and signalling correlate with increased plant resistance against various pathogens. However, the underlying molecular mechanisms whereby CK signalling modulates plant immune networks are not fully understood. To investigate mechanisms how CK responses culminate in protecting plants from infection with model pathogen Pst DC3000, we mapped high quality Arabidopsis Protein-Protein Interactions (Y2H-PPI). A dynamic of change to the basal network is given by transcriptome data after treatment of Arabidopsis with Pst DC3000. Nodes of the cellular interactome enriched in immune functions and their interacting partners are organized into sub-networks. According to our analysis, seventeen proteins of CK canonical pathway are involved in immune network. Based on topological analyses, nodes connectivity and network modularity, we identified functional hubs in our sub-networks. Two of the CK signalling proteins AHK2 (receptor) and ARR14 (type-B response regulator) are among the top ten most connected hubs. Besides, we also conducted analyses on modulated CKs conditions such as deletion of type-A ARRs as well as exogenous CK application. These analyses culminate in a functional core module where nodes of SA mediated defense and Cytokinins pathway actively participate. Furthermore, we established a Boolean model of plant immune and pathogen virulence specific proteins. Model analyses and dynamic simulations for various hormone types with and without pathogen infection manifested diverse immune responses. Taken together, our integrated systems biology approaches are highly instrumental in deciphering hormonal crosstalk in plant immunity.

1

IL-1

mailto:[email protected]

3

Induced auxin overexpression reveals a link between auxin signaling and lignification through the induction of ethylene biosynthesis and several cell wall modifying enzymes

Marta-Marina Pérez Alonso, Daniel Jesús García Martínez, Stephan Pollmann* Centro de Biotecnología y Genómica de Plantas (UPM – INIA), Campus de Montegancedo, 28223 Pozuelo de Alarcón (Madrid), Autovía M-40 km 38, Spain * corresponding author ([email protected])

Auxin is associated with the regulation of virtually every aspect of plant growth and development. Many previous genetic and biochemical studies revealed that, among the proposed routes for the production of auxin, the indole-3-pyruvic acid (IPyA) pathway is the main source for indole-3-acetic acid (IAA) in plants. The IPyA pathway involves the action of 2 classes of enzymes, tryptophan-pyruvate aminotransferases (TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (TAA 1)/TRYPTOPHAN AMINOTRANSFERASE RELATED (TAR)) and flavin monooxygenases (YUCCA). Both enzyme classes appear to be encoded by small gene families in Arabidopsis consisting of 5 and 11 members, respectively. We recently showed that it is possible to induce transcript accumulation of 2 YUCCA genes, YUC8 and YUC9, by methyl jasmonate treatment (Hentrich et al. 2013a). Both gene products were demonstrated to contribute to auxin biosynthesis in planta. Furthermore, we reported that the overexpression of YUC8 as well as YUC9 led to strong lignification of plant aerial tissues likely due to the up-regulation of ethylene biosynthesis (Hentrich et al. 2013b). New transcriptomic and metabolomic evidence indicates that this abnormally strong secondary growth is not only linked to increased levels of ethylene production, but also to substantial changes in the expression level of a subclass of cell wall modulating enzymes. Those enzymes play a role in cell wall softening and consequently in the modification of the secondary cell wall composition. Hentrich M., Böttcher C., Düchting P., Cheng Y., Zhao Y., Berkowitz O., Masle J., Medina J., Pollmann S. (2013a). Plant Journal 74 (4): 626-637. Hentrich M., Sánchez-Parra B., Pérez Alonso M. M., Carrasco Loba V., Carrillo L., Vicente Carbajosa J., Medina J., Pollmann S. (2013b). Plant Signaling & Behavior 8 (11): e26363.

1

IL-2


4

A systems biology approach to the evolution of virus-plant interactions

Santiago F. Elena* Instituto de Biología Molecular y Celular de Plantas (CSIC-UPV), Campus UPV CPI 8E, Ingeniero Fausto Elio s/n, 46022 València, Spain; The Santa Fe Institute, 1399 Hyde Park Road, Santa Fe, NM 87501, USA * corresponding author ([email protected])

Understanding the mechanisms by which viruses overcome host defenses in order to proliferate has been a challenging problem owing to the multiplicity of factors and complexity of interactions involved. The advent of genomic techniques has opened the possibility to grasp a global picture of the interaction. We used Arabidopsis thaliana to identify and compare genes that are differentially regulated upon infection with seven distinct plant viruses (Rodrigo et al. 2012). In a first approach, we established lists of genes differentially affected by each virus and compared their involvement in biological functions and metabolic processes. We found that phylogenetically-related viruses significantly alter the expression of similar genes and that viruses that naturally infect Brassicaceae display a greater overlap in the lists of altered genes. In a second approach, virus-manipulated genes were contextualized using transcriptional and protein-protein interaction networks of A. thaliana. This method uncovered a general mode of action of plant viruses, in which perturbations preferentially affect genes that are highly connected, central and organized in modules. Rodrigo G., Carrera J., Ruiz-Ferrer V., Del Toro F. J., Llave C., Voinnet O., Elena, S. F. (2012). PLoS ONE 7, e40526.

1

IL-3


5

Digging potato molecular responses to virus infection

Špela Baebler* National Institute of Biology, Večna pot 111, Ljubljana, Slovenia *corresponding author ([email protected])

Potato (Solanum tuberosum L.) is most widely grown tuber crop and the fourth most important food crop with a continuous progression of the world production of tubers that is challenged by changing environment and new emerging diseases. Understanding its biology by using systems biology tools can contribute to plant breeding and development of efficient agricultural practices. However, the exploitation of data obtained by high-throughput wet lab methods is hindered by dispersed data and unavailability of potato specific functional annotation and visualization tools. The genome of diploid potato was sequenced three years ago and enabled better integration of the existing data. To combine existing information on potato genes and enable data integration from various datasets, we have merged information from two potato gene models and two sets of potato unigens to 35609 gene paralog groups. For easier data interpretation we have adapted plant-specific MapMan ontology and designed a centrally maintained database and a web interface GoMapMan (www.gompaman.org, Ramšak et al., 2013) for browsing, searching and editing the ontology and gene annotations. Within the ontology, genes of the model species Arabidopsis and 4 crop species (potato, tomato, tobacco and rice) are connected trough ortholog groups. Developed tools have allowed us translation of existing knowledge on model and other crop species to potato as well as better visualization and interpretation of high-throughput experimental data on potato - Potato virus Y interaction. Ramšak Ž., Baebler Š., Rotter A., Korbar M., Mozetič I., Usadel B., Gruden K. (2014). Nucleic acids research 42, D1167–75.

1

IL-4

mailto:[email protected]://www.gompaman.org/

6

Sukkula retrotransposon insertion polymorphisms in barley calli

Nermin Gozukirmizi*, Gonul Kartal-Alacam, Sibel Yilmaz, Sevgi Marakli Istanbul University, Faculty of Science, Department of Molecular Biology and Genetics, 34134, Vezneciler, Istanbul-TURKEY *corresponding author ([email protected])

Barley has economical value and it is an important model plant for genetic studies. Transposons cover more than 80% of barley genome. More than 40 retrotransposons were characterized in barley genome. Some of them are autonomous but some are not. This type of transposons replicates via RNA and moves in the genome. As a result of these movements, mutations and genome enlargements are occurred. Somaclonal variations are spontaneously occurred variations in tissue culture conditions. These variations could be produced by genetic and/or epigenetic mechanisms and result in problems in gen transfer applications. In this study, mature barley embryos (Hordeum vulgare cv. Tokak 157/37) were cultured for callus formation. Sukkula (a non-autonomous retrotransposon) polymorphism was investigated in calli with different culturing time (40 and 80 day-old) which was derived from the same embryo in MS medium supplemented with 3 mg/l Dicamba. Polymorphisms were detected using Inter-Retrotransposon Amplified Polymorphism (IRAP) and Inter Primer Binding Site Amplification (iPBS) techniques with two different Sukkula-specific primers. Non-cultured 5 mature embryos were used as control. While up to 14% polymorphism rates were calculated from non-cultured embryos, 61% and 0% polymorphism rates were observed with IRAP analyses in 40 and 80 day-old calli, respectively. iPBS analyses revealed polymorphism ratio up to 29% in non-cultured materials, and up to 58% and 70% in 40 and 80 day-old calli materials, respectively. These findings show that tissue culture conditions and culturing time cause insertional activation of some barley retrotransposons. These stress induced activation might be responsible for development of somaclonal variations.

1

SL-1


7

Suitability of wild potato relatives for functional analysis of potato genes using virus induced gene silencing

David Dobnik1,*, Maja Jamnik1, Vivianne G.A.A. Vleeshouwers2, Kristina Gruden1, Jana Žel1 1 Department of biotechnology and systems biology, National institute of biology, Večna pot 111, 1000 Ljubljana, Slovenia 2 Wageningen UR Plant Breeding, Wageningen University and Research Centre, P.O. Box 386, 6700 AJ, Wageningen, The Netherlands * corresponding author ([email protected])

Potato is the fourth most important food crop worldwide. It is susceptible to many pathogens, where potato virus Y (PVY) is one of the most important ones. To better understand the interaction between potato and PVY several “omics” studies were already performed, resulting in a plethora of interesting genes. However, when it comes to the confirmation of importance of individual genes by the means of functional analysis, the results are rather scarce, as the cultivated potatoes are not the optimal model organisms. To provide the scientific community with the most suitable system for functional analyses of genes from potato using virus induced gene silencing (VIGS), we have screened large panel of wild potato relatives for their susceptibility to VIGS. Based on a visual marker (silencing of PDS) we have selected 15 appropriate Solanum species for further evaluation. We have challenged the selected wild potato relatives with PVY and followed the spreading of the virus. To check whether the empty silencing vector (based on Tobacco rattle virus) has an effect on PVY spreading, we performed VIGS with empty vector and then inoculated the same plants with PVY. Finally we challenged the selected wild potato relatives with PVY labelled with GFP (PVY-GFP). Based on the results of these experiments the most appropriate candidate for VIGS experiments was selected. To confirm that the selected species is suitable for VIGS studies of potato genes, we have silenced some of the selected genes from potato and observed the effect on the viral spreading with the use of PVY-GFP. Our results have shown that the selected system is appropriate for functional analysis of genes from potato.

1

SL-2


8

Functional analysis of an Ethylene Response Factor involved in potato defence response to PVY

Anna Coll*, Ana Lazar, David Dobnik, Katja Stare, Špela Baebler, Kristina Gruden

Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia * corresponding author ([email protected])

Potato (Solanum tuberosum L.) is the world’s most widely grown tuber crop and potato virus Y (PVY) is one of the major potato pathogen causing severe crop loss in different areas worldwide. To better understand the potato defence response against PVY we studied the role of ethylene response factor (ERF) genes since they function as important plant-specific transcription factors in regulating biotic and abiotic stress response. A potato ERF gene, StERF, that was identified and selected based on previous transcriptomics experiments performed in our group has been characterised and functionally analysed in this study. Expression patterns of the gene in hypersensitive resistance (HR) potato cultivar infected with PVY pointed to its importance as a signalling component in potato defence response. Localisation studies showed that StERF strongly accumulated in cell nucleus after PVY infection. In addition, using virus-induced gene silencing (VIGS) we demonstrated that PVY systemic spread is delayed in StERF silenced plants. We further examined the potential hormonal signalling involved in the expression of StERF gene and demonstrated that our gene integrates several signalling pathways. Taken together our results suggested the importance of SotubERF in potato defence mechanisms against PVY and its involvement in the ethylene signalling pathway. Therefore the data contributes to better understand the complex network of plant defence signalling pathways.

1

SL-3


9

Involvement of potato (Solanum tuberosum L.) MKK6 in response to Potato virus Y

Ana Lazar*, Anna Coll, David Dobnik, Špela Baebler, Jana Žel, Kristina Gruden Department of biotechnology and systems biology, National institute of biology, Ljubljana, Slovenia * corresponding author ([email protected])

Mitogen-activated protein kinase (MAPK) cascades have crucial roles in the regulation of plant development and in plant responses to stress. Plant recognition of pathogen-associated molecular patterns or pathogen-derived effector proteins has been shown to trigger activation of several MAPKs. This then controls defence responses, including synthesis and/or signalling of defence hormones and activation of defence related genes. The MAPK cascade genes are highly complex and interconnected, and thus the precise signalling mechanisms in specific plant–pathogen interactions are still not known. Here we investigated the MAPK signalling network involved in immune responses of potato (Solanum tuberosum L.) to Potato virus Y, an important potato pathogen worldwide. Sequence analysis was performed to identify the complete MAPK kinase (MKK) family in potato, and to identify those regulated in the hypersensitive resistance response to Potato virus Y infection. Arabidopsis has 10 MKK family members, of which we identified five in potato and tomato (Solanum lycopersicum L.), and eight in Nicotiana benthamiana. Among these, StMKK6 is the most strongly regulated gene in response to Potato virus Y. The salicylic acid treatment revealed that StMKK6 is regulated by the hormone which is in agreement with the salicylic acid-regulated domains found in the StMKK6 promoter. The involvement of StMKK6 in potato defence response was confirmed by localisation studies, where StMKK6 accumulated strongly only in Potato-virus-Y-infected plants, and almost exclusively in the cell nucleus. Using a yeast two-hybrid method, we identified three StMKK6 targets downstream in the MAPK cascade: StMAPK4_2, StMAPK6 and StMAPK13. These data together provide further insight into the StMKK6 signalling module and its involvement in plant defence.

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Using a magnetic particle system in plant NA purification

Suomalainen Sini*

Thermo Fisher Scientific, Vantaa, Finland * corresponding author ([email protected])

Sample preparation is often a limiting step for different kind of genomics studies. By using magnetic particles for nucleic acid isolation, it is possible to speed up the purification process and achieve better results from variety of sample types. In this presentation there will be an overview to the magnetic particle purification and examples how to enhance the process by using KingFisher magnetic particle system. The KingFisher system is an open platform allowing the user to choose any available magnetic particle purification reagents from different manufacturers. There are instruments available for low throughput research laboratories as well as routine testing laboratories needing higher throughput. The high-throughput extraction of plant nucleic acids is very important for several applications like the plant breeding, quality control of seeds and other molecular screening tests of plants. With the KingFisher system, highly pure DNA can be obtained from fresh, frozen, dried or lyophilized plant samples. Magnetic particle purification is not only efficient and reproducible, but also a gentle way to isolate good quality nucleic acids, also intact RNA from fresh and frozen plant samples.

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Genome-specific distinction between Brassica oleracea L. and Sinapis arvensis L. seeds in seed lots

Barbara Pipan*, Romana Rutar, Darja Vouk, Vladimir Meglič Agricultural institute of Slovenia, Hacquetova ulica 17, SI-1000 Ljubljana * corresponding author ([email protected])

Diverse Brassicaceae family includes economically important Brassica L., Sinapis L. and Raphanus L. species. Brassica oleracea L. (CC genome) and Sinapis arvensis L. (SS genome; also known as Brassica kaber (DC.) L.C. Wheeler) are closely related diploid species (2n=18). B. oleracea is widely used as vegetable crop; S. arvensis is mostly appeared as weedy/wild plant with a series of superior traits, since is becoming an important germplasm resource in Brassicaceae breeding schemes. Close relationsip between B. oleracea and S. arvensis is reflected in a high level of morphological, genetic and karyotypic similarity. Consequently, it is difficult to distinguish these two species, especially when analyzing seed purity in seed testing laboratories. The problem lies primarily in seed identification, where seeds of S. arvensis represent impurities in B. oleracea seed lots. The method to distinguish between these two species according to the seed coat morphology, using binocular at low magnification (x40), is not reliable. The fast and cost effective method is required to provide accurate information for species identification. Molecular markers, such as SSRs (Short Sequence Repeats), could be readily applied due to their informative nature.

To distinguish between B. oleracea and S. arvensis species on molecular level, we developed fast and reliable SSR method using genome-specific markers. According to Pipan et al. (2013), we tested 15 highly informative SSR markers, originating from different Brassica species. DNA was extracted from germinated seedlings of morphologically identified S. arvensis or B. oleracea seeds used for a seed purity test. Reference DNA (internal resource) from both observed species was included in the study. Two markers (Na12-C08 and Ra2-E12) were proven to be genome-specific due to their successful amplification in B. oleracea only, but not in any of the S. arvensis samples. The third marker (Ra2-A01) was amplified in all samples, but in a different allele forms that distinguished B. oleracea (at ~ 150 bp length) and S. arvensis (at ~ 180 bp length) genomes. Moreover, this locus could also be applied for intra-specific differentiation within S. arvensis genome.

The use of genome-specific markers (Na12-C08, Ra2-E12 and Ra2-A01) in seed testing laboratories would provide a fast, reliable and cost-effective tool for qualitative genetic screening and identification of S. arvensis seeds as impurities in B. oleracea seed lots or vice versa.

Pipan B., Šuštar-Vozlič J., Meglič, V. (2013). Genetika 45 (2), 309-327.

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Plant pathogens detection by simultaneous multiplex isothermal amplification and detection of RNA and DNA targets

David Dobnik*1, Dany Morisset1,2, Rok Lenarčič1, Maja Ravnikar1 1 Department of biotechnology and systems biology, National institute of biology, Večna pot 111, 1000 Ljubljana, Slovenia 2 Current address: CropDesign N.V., Technologiepark 21C, 9052 Gent (Zwijnaarde), Belgium * corresponding author ([email protected])

The impact of climate change and expansion of trade networks is expected to cause an emergence of new plant pathogens in places, where they have not been present before. Reliance on sometimes lengthy laboratory testing can cause significant delays in preventive actions. A relatively small number of detection tools is currently available for field use with many having serious shortcomings in terms of sensitivity, complexity or cost. Thus, there is a real need for rapid, simple and robust detection methods that can be used in the field to allow early and simultaneously detection of several plant pathogens. Molecular detection methods for plant pathogens mostly rely on amplification of specific genetic elements. To reduce the costs and time needed for analyses, the development should be focused to detection methods allowing multiplex amplification and detection. The genetic material of pathogens can be carried in RNA or DNA form, therefore the method for their multiplex detection should also enable amplification and detection of both in parallel. We developed an isothermal nucleic-acid-sequence–based amplification [NASBA]-implemented microarray analysis (NAIMA) suitable for simultaneous multiplex amplification of RNA and DNA targets and their detection on ArrayTubes™. The method was demonstrated to be very sensitive and specific for detection of two economically important quarantine plant pathogens, Potato Spindle Tuber Viroid (RNA target) and Ralstonia solanacearum (DNA target). The advantage of the NAIMA method is that it is isothermal and therefore no expensive apparatus is needed for work, which makes it also suitable for the determination of plant pathogens directly in the field. The method could also be applicable to any domain where there is need to detect RNA and DNA pathogens or other targets simultaneously.

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Interactions between components of the plant defence response and viral proteins

Živa Ramšak*1, Tjaša Stare1, Tjaša Lukan1, Dragana Miljković2, Kristina Gruden1

1 Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia 2 Department of Knowledge Technologies, Jožef Stefan Institute, Ljubljana, Slovenia * corresponding author ([email protected])

Infection of a plant by a pathogen initiates a complex interaction between both players involved, leading to changes in the complex signalling network, which result in gene activity changes and reprogramming of the cell metabolism. A systems biology approach was adopted for the purpose of modelling complex biological processes in order to understand the mechanisms and dynamics involved in potato plant defense following the infection with potato virus Y (PVY).

A qualitative model of potato plant defence signalling network (PDS) was constructed (Miljkovic et al., 2012), describing the biosynthesis and signal transduction pathways for three crucial phytohormones involved in plant defence: salicylic acid (SA), jasmonic acid (JA) and ethylene (ET). The prior knowledge from literature was expanded with information on the viral and plant component interactions, protein-protein interactions and protein-DNA interactions in plant Arabidopsis.

The resulting robust qualitative model offers new insights into the plant-virus interaction by expanding the knowledge on critical components of plant defence signalling, thus producing novel hypotheses to be tested in the wet lab. First efforts of dynamical modelling of the selected ethylene model sub-component were already performed, indicating the importance of protein degradation efficiency to obtain biologically relevant representations of network responsiveness.

Miljkovic, D., Stare, T., Mozetic, I., Podpecan, V., Petek, M., Witek, K., Dermastia, M., Lavrac, N., Gruden, K. (2012). PloS ONE, 7(12), e51822. doi:10.1371/journal.pone.0051822

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2 A Interakcije rastlin z okoljem in drugimi organizmi

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Dynamic compartment specific changes in glutathione and ascorbate levels in plants under different environmental conditions

Maria Müller*, Bernd Zechmann Institute of Plant Sciences, University of Graz, Schubertstraße 51, Graz, Austria * corresponding author ([email protected])

Ascobate and glutathione are the most important antioxidants in plants. They are involved in the

detoxification of reactive oxygen species (ROS), redox signaling, in the modulation of defense gene

expression and they are important for the regulation of enzymatic activities. For these reasons levels of

ascorbate and glutathione are often used as stress markers in plants. Nevertheless, most studies in the

past focused on measuring ascorbate and glutathione in whole tissues and organs rather than in single

cells and organelles. Considering that stress situations affect cell compartments differently and that

ascorbate and glutathione metabolism is highly compartment specific we applied this method on different

plant species during abiotic and biotic stress situations to gain thorough knowledge about the subcellular

distribution of ascorbate and glutathione in plants and on the importance of these antioxidants in certain

cell compartments during stress situations.

Acknowledgement: This work was supported by the Austrian Science Fund.

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Abiotic stress in agricultural plants: Phaseolus sp. and drought

Jelka Šuštar-Vozlič1*, Marko Maras1, Tanja Zadražnik1, Marjetka Kidrič2, Mateja Zupin1, Jaka Razinger1, Dominik Vodnik3, Vladimir Meglič1

1 Agricultural Institute of Slovenia, Hacquetova ulica 17, 1000 Ljubljana Slovenia 2 Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia 3 Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia * corresponding author ([email protected])

Different abiotic factors such as drought, flood, extreme temperatures, high salinity and others affect plant growth and crop productivity. Drought contributes to considerable yield losses in different crops, including common bean (Phaseolus vulgaris L.). The response of common bean to drought is not well characterized yet. The aim of our research has therefore been oriented towards obtaining a deeper insight into the molecular and morpho-physiological adaptation of bean plants to drought. Differential gene expression study was conducted on leaves of eight common bean genotypes at different levels of dehydration. The results revealed the alteration of 15 drought-responsive transcripts. The expression profiles of selected genes were studied also in different bean species. The mode of expression was found consistent within Phaseolus spp. A proteomic approach was used to identify drought-responsive proteins in leaves of two cultivars differing in the response to drought, Tiber and more sensitive Starozagorski čern. The identified proteins were divided into different functional categories that include metabolic proteins, cellular defence/stress proteins, proteins involved in reactions of photosynthesis and proteins with unknown function. Analysis of stem proteome of cultivar Tiber under drought stress using in-gel stable isotope labelling revealed proteins grouped into several functional groups. By studying proteases we have shown that there are several serine endopeptidases and aminopeptidases in leaves of common bean which are differently affected by drought. A protease from leaves of cultivar Zorin has been studied in details; characterization at the protein and gene level led to its classification as a new plant subtilisin-like serine protease, named PvSLP2. Genetic characterization of cvs. Tiber and Starozagorski čern and 82 recombinant inbred lines (RILs) using molecular markers has been launched recently. The main aim is to establish genetic map of bean genome, which will be used in identification of markers linked to the quantitative trait loci involved in drought response of common bean. Several hundred microsatellites have been included in the characterization and different physiological parameters were measured in parental cultivars and RILs in order to define differences of their response to drought. The integrated knowledge of the genomic, proteomic and physiological processes during drought response will contribute to the basic understanding of drought tolerance, not only in bean but also in other plants, and will offer novel opportunities for bean management in different environments.

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Risk assessment of receptor organisms: analysis of intake of metals through food chain

Samar Al Sayegh Petkovšek1*, Nataša Kopušar1, Davorin Tome2, Boris Kryštufek3 1 ERICo Velenje, Inštitut za ekološke raziskave d.o.o., Koroška 58, 3320 Velenje 2 Nacionalni inštitut za biologijo, Večna pot 111, 1000 Ljubljana 3 Prirodoslovni muzej, Prešernova 20, 1000 Ljubljana * corresponding author ([email protected])

On the basis of the results for heavy metal levels in soil, in herbs and in tissues of receptor organisms (small mammal species, passerine birds) obtained in the period 2011-2014 in differently polluted areas in Slovenia, the following conclusions can be made. (a) The soil in the vicinity of the former smelter (Žerjav) is heavily polluted with Pb, Cd, Zn and As, and burdened with Mo and Hg. The latter is reflected in increased levels of metals in herbs (Cd, Pb, Zn, Mo, Ni) and in earthworms (Pb, Cd, Zn). Particularly problematic is Cd, which accumulates in herbs (Sesleria albicans Kit. Ex Schultes) such as in earthworms. Pb accumulates only in earthworms, but the intake of this element in the herbs is quite large, despite its relatively poor mobility (BCF: 0.08 to 0.50), due to the extremely high levels of Pb in soil. Therefore, these two dietary sources (herbs, earthworms) could be the most significant route of Pb and Cd exposure for receptor organisms. To a much lesser extent this applies to fruits and seeds of tree and shrubs species (Cornus sanguinea L., Sorbus aria (L.) Crantz, Acer pseudoplatanus L. and Pinus sylvestris L.). The mean Pb and Cd concentrations in liver of small mammals species (Microtus agrestis (Linnaeus, 1761)), Myodes glareolus (Schreber, 1780), Apodemus flavicollis (Melchior, 1834)), reached effect concentrations in a significant proportion of the livers of the small mammal specimens (Pb: 40%, Cd: 67%). These findings indicate that the majority of small mammals trapped in the area of the former lead smelter are at risk of toxicological effects due to the very high uptake of Pb and Cd in the organism. (b) On the contrary, Pd and Cd concentrations in livers of small mammals from the other study areas (vicinity of the largest Slovenian thermal power plant (Veliki Vrh) and along the main road (Črnova)) were comparable with reference values and considerably lower than effect concentrations. The smallest levels of pollutants in all types of samples were found in soil, plant and animal samples from reference location (The Logar Valley). In general, these differences were statistically significant. (c) Pb and Hg in the tail feathers of passerine birds from polluted study areas were statistically significant higher in comparison with reference area. (d) Hazard Quotient (HQ) higher than 1 (existence of risk) was calculated for Myodes glareolus, sampled in the vicinity of the lead smelter, if the proportion of the roots in its diet is at least 20% and for Apodemus flavicollis, collected from the same study area, if the proportion of the earthworms in its diet is at least 10%.

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Biomonitoring of temporal and spatial patterns of metal deposition in Slovenia during the last twenty years

Zvonka Jeran1*, Darja Mazej1, Mitja Skudnik2

1Department of Enivironmental Sciences, “Jožef Stefan” Institute, Jamova 39, 1000 Ljubljana, Slovenia 2Slovenian Forestry Institute, Večna pot 2, 1000 Ljubljana, Slovenia

* corresponding author ([email protected])

Several country wide monitoring surveys were performed since 1992 in Slovenia in which two types of biomonitoring organisms were used to assess metal and nitrogen air pollution. In 1992 and 2001 epiphytic lichen Hypogymnia physodes was collected and analysed for more than 20 elements, however in 1995 also moss Hypnum cupressiforme was for the first time used in our country as a monitor organism as a part of European Moss Survey within the UN-ECE ICP-Vegetation. Moss monitoring was since then regularly repeated at 5 years intervals. Moss samples were always collected in the period between the middle of June and the end of August at the regular grid of 16 x 16 km or 16 x 8 km (2010), though the number of plots at which samples were taken differed among the surveys, namely from 30 in 1995 to 102 plots in 2010, of which 32 plots were the same in all the last three surveys. Over the 15 years there was also a change in methods for elemental determination, namely since 2005 ICP-MS was used instead of Instrumental neutron activation analysis. All together 17 elements were determined: As, Cd, Cr, Co, Cu, Fe, Mo, Ni, Pb, Sb, Se, Sr, V, Ti and Zn. For Hg determination cold vapour AAS (2005) or direct measurement using the DMA method (2010) was used. Also since 2001 nitrogen and sulphur were analysed using the CNS method. Although there were slight modifications in moss sampling over the 15 year period, the spatial pattern of elements and their temporal trends well reflect the overall pollution in the country, as well as also the natural background. Similarly to the majority of European countries, a decrease of element concentrations was observed in Slovenia since 1995, which was most probably due to economic factors since some of the heavy polluters have shut down or modernized their technology to decrease emissions. Another reason for the great decline as observed in median levels of all elements on a national scale in 2010 in comparison to 2005 might lie in the fact that in 2010 special attention was devoted to sampling and testing the influence of environmental parameters on element levels in mosses. The spatial pattern of levels of anthropogenically derived elements especially Pb, Cd, Mo, Zn, Sb and nitrogen in Slovenia can be explained on the one hand by the population density and the locations of the main pollution sources and on the other hand, by transboundary air transport of pollutants from neighbouring countries, mainly as a consequence of meteorological conditions (wind, precipitation).

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Soil abiotic factors determine arbuscular mycorrhizal fungal communities in hypoxic soils at natural CO2 springs (mofettes)

Irena Maček1*, Nataša Šibanc1, Alex J. Dumbrell2, Thorunn Helgason3 1 Department of Agronomy, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1001 Ljubljana, Slovenia 2 School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK 3 Department od Biology, Wentworth Way, University of York, York YO10 5DD, UK * corresponding author ([email protected])

Natural CO2 springs (mofettes) are specific and extreme ecosystems, characterized by high soil CO2 and reduced soil O2 concentrations. A locally hypoxic environment is established that is similar to those found in waterlogged or flooded soils. In mofettes, geological CO2 exhalations result in a long-term abiotic selection pressure affecting local soil biota. The characterization of arbuscular mycorrhizal (AM) fungal communities from plant roots sampled in the Stavešinci mofette area (Slovenia), confirmed the presence of apparently unique AM fungal assemblages across a range of soil CO2 concentrations. We examined whether AM fungal taxa abundant in locations with high soil CO2 concentrations are local specialists or widespread generalists able to tolerate a range of different soil conditions. To answer this, a high throughput sequencing approach (Roche 454 sequencing) was used to see if those taxa present in areas with high soil CO2 concentrations are also found in low abundance among AM fungal communities from soils with ambient CO2 concentrations. In addition to hypoxia, we investigated other abiotic soil factors (e.g. soil pH) driving changes in AM fungal communities under the extreme environmental conditions found in mofette sites. Results show that some fungi are more strongly associated with local variations in the soil environment, particularly hypoxia, than with other abiotic soil characteristics or distributions of their host plants.

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Physiological studies of drought stress in hop (Humulus lupulus L.)

Rozalija Povše1*, Dominik Vodnik2, Stanislav Mandelc2, Branka Javornik2, Andreja Čerenak1 1 Inštitut za hmeljarstvo in pivovarstvo Slovenije, Cesta Žalskega tabora 2, 3310 Žalec 2 Biotehniška fakulteta, Jamnikarjeva 101, 1000 Ljubljana * corresponding author ([email protected])

Frequent droughts, irregular precipitation patterns and limited sources of water dictate plant breeding programmes oriented to the selection of crops with higher tolerance to water stress. Combined use of omic approaches (transcriptomics, proteomics) and conventional physiological techniques (gas exchange techniques, fluorescence, water potential measurements) enables a better insight into the response of plants, which is crucial for selecting targeted properties of drought tolerant cultivars. This approach was used in a study with two hop cultivars, Aurora (tolerant) and Savinjski golding (susceptible). Two pot experiments were run in two consecutive seasons. Measurements and sampling of leaves were done at different stages of developing drought, documented by soil water content, relative water content in leaves and leaf water potential measurement. In our contribution we will present results on stomatal and non-stomatal inhibition of photosynthesis, differences in water use efficiency and photochemical efficiency of the tested cultivars, and the preliminary results of differential proteomics.

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UV radiation and selenium addition affects silicon accumulation in buckwheat plants

Aleksandra Golob1*, Vekoslava Stibilj2, Ivan Kreft3, Mateja Germ1 1 Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia 2 Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenija 3 Slovenian Forestry Institute, Večna pot 2, SI-1000 Ljubljana, Slovenia *corresponding author ([email protected])

UV radiation affects organisms, especially plants, on different levels. UV radiation is consequently a carrier of information about the environment of plants. However, when exposed to enhanced doses of UV radiation, plants can be damaged because of the enhanced production of reactive oxygen species which cause oxidative stress to plants. Selenium (Se) is one of biologically essential elements for animals and humans, needed for activity of antioxidative enzymes, such as glutathione peroxidase and thioredoxin reductase. Se has not been classified as an essential element for plants, but it has been considered as beneficial. It has been demonstrated that Se can increase the tolerance of plants to UV-induced oxidative stress and promotes the growth of plants subjected to high-energy radiation. In higher concentrations Se has toxic effect. Silicon (Si) is second most abundant element in Earth crust. Si has an important role in stress mitigation, through increased production of antioxidants and its binding to, and co-precipiataion with metal ions. We investigated the effect of different levels of UV radiation and foliar spraying with Se on biochemical and physiological traits as well as Si accumulation in leaves of Fagopyrum tataricum and Fagopyrum hybridum. Experimental plants were exposed outdoors to four treatments regarding the quantity of UV radiation (ambient or reduced) and added Se (naturally accessible or foliarly treated with Na selenate). The content of pigments and UV absorbing compounds, transpiration rate, photochemical efficiency of PS II, respiratory potential and the biomass of plant parts were measured. UV treatment had no effect on pigment content, but enhanced potential photochemical efficiency at the early stage of development of both buckwheat species. Addition of Se increased effective photochemical efficiency and transpiration, while decreased potential photochemical efficiency. In condition of reduced UV radiation addition of Se decrease respiratory potential, while increased it under ambient UV conditions. Just the opposite effect was observed for the mass of the seeds. Preliminary results showed that addition of Se significantly increased accumulation of Si in leaves of both buckwheat species. In Tartary buckwheat UV radiation had positive effect on Si accumulation, while in hybrid buckwheat there were no differences between UV treatments.

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Growth in conifer and deciduous trees under enhanced UV-B radiation

Tadeja Trošt Sedej1*, Dušan Rupar1, Alenka Gaberščik1 1

Department of Biology, Biotehnical Faculty, University of Ljubljana, Večna pot 111, 1000 Ljubljana, Slovenia

* corresponding author ([email protected])

Trees, as plants in general, possess diverse systems which respond to UV-B radiation. The results available in the literature for conifer and deciduous trees indicate that UV-B radiation may affect those tree groups differently according to their physiology, biochemistry and morphology, to needle and leaf development stage, to interaction with environmental conditions and multilevel interactions between all those factors. We compared strategies in Norway spruce (Picea abies (L.) Karst.) and European beech (Fagus sylvatica L.) for coping with enhanced UV-B radiation. The two tree species were exposed outdoors to enhanced UV-B for three years during which time, selected parameters were monitored (Trošt Sedej & Gaberščik 2008, Trošt Sedej & Rupar 2013). Spruce and beech exhibited great variability in the amounts of chlorophyll, methanol-soluble UV-B and UV-A absorbing compounds, and optimum quantum yield of photosystem II. The effects of UV-B radiation varied with needle and leaf development stage and interaction with environmental conditions. Enhanced UV-B radiation triggered responses in both trees and a reduced negative effect of UV-B radiation on spruce photochemical efficiency was observed during prolonged drought. The results show high UV-B tolerance of both tree species and indicate the complexity of plant response to UV-B, involving multilevel interactions with environmental factors. Trošt Sedej, T., Gaberščik, A. (2008). The effects of enhanced UV-B radiation on physiological activity and growth of Norway spruce planted outdoors over 5 years. Trees 22 (4), 423-435. Trošt Sedej, T., Rupar, D. (20013). Deciduous and evergreen tree responses to enhanced UV-B treatment during three years. Acta Biologica Slovenica 56 (2) 35-50.

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The functioning of two karst ecosystems revealed by eddy-covariance measurements

Klemen Eler1*, Mitja Ferlan2, Giorgio Alberti3, Alessandro PERESSOTTI3, Primož Simončič2, Dominik Vodnik1 1 University of Ljubljana, Biotechnical Faculty, Jamnikarjeva 101, Ljubljana, Slovenia 2 Slovenian Forestry Institute, Večna pot 2, Ljubljana, Slovenia 3 Department of Agricultural and Environmental Sciences, University of Udine, Udine, Italy * corresponding author ([email protected])

Since 1980's the eddy covariance measurements of trace gasses, latent and sensible heat have made their way in ecosystem research, providing many valuable information on yearly, seasonal and daily cumulatives of carbon (C) and water vapour exchange of different ecosystems of the world. Moreover, this key ecosystem research method greatly increased our understanding of environmental regulation of carbon emission and consumption rates which is inevitable input information to predict global change effects of different ecosystems and to designate a proper management. In this contribution carbon and water vapour exchanges of a spatially heterogeneous tree-shrub-grassland mosaic developed on former semi-dry calcareous grassland in the sub-Mediterranean region of Slovenia were studied and compared with the nearby grassland still in use as a pasture. Yearly and seasonal net ecosystem exchange and evapotranspiration of both ecosystems were examined together with their environmental controls (soil water content, air temperature, light availability) during the period August 2008 - December 2012. Results showed significant differences in yearly cumulatives, seasonal phenology of C assimilation and drought resistance between the investigated ecosystems. The occurrence of drought seemed to largely govern ecosystem functioning and productivity. By identifying the drought periods objectively using the evapotranspiration response at different soil water content values it was shown that more than the severity of drought the duration of the longest yearly drought affected the estimated yearly gross primary productivity. The functioning of ecosystems was also compared with respect to ecosystem-level water use efficiency, rain use efficiency and light use efficiency in different environmental conditions and phenological phase.

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Change of phenolic compounds pattern in Nicotiana megalosiphon as a response to abiotic and biotic stress

Saša Likić*, Gordana Rusak University of Zagreb, Faculty of Science, Department of Biology, Marulicev trg 9a, 10000 Zagreb, Croatia * corresponding author ([email protected])

Quantitative and qualitative changes of phenolic compounds were compared in untreated, wounded, Cucumber mosaic virus (CMV) infected and in Cucumber mosaic virus containing satellite RNA (CMVsat) infected Nicotiana megalosiphon plants. Significant changes in total phenolics (TP) content were analysed at five different time intervals: immediately after inoculation, 2, 4, 7 and 14 days after inoculation. The highest decrease of TP was detected in wounded while the highest increase of TP was detected in both CMV and CMVsat infected plants. Defence response to wounding in investigated plants resulted in a significant decrease of detected phenolics content compared to untreated plants. On the other hand, significant increase in the content of detected phenolics in infected plants compared to wounded plants was detected. Nevertheless, the content od phenolics in CMVsat infected plants was significantly higher compared to the content in CMV infected plants. Depending on treatment and on time interval, significant increase or decrease of quercetin or kaempferol was detected in investigated species. Observed changes of phenolics suggest distinct responses to wounding and virus infection, which could probably be involved in activating different defence response to a specific stress.

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Phenols, photosynthetic pigments and leaf morphology as biomarkers of environmental influences on Posidonia oceanica (L.) Delile

Željka Vidaković-Cifrek1*, Mirta Tkalec1, Jasna Dolenc Koce2, Donat Petricioli3, Adam Cvrtila1, Jelena Katanović1, Janja Maltar1, Mateja Smiljanec1, Nina Zupanič2, Tatjana Bakran-Petricioli1 1University of Zagreb, Faculty of Science, Department of Biology, Rooseveltov trg 6, HR-10000 Zagreb, Croatia; 2University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, SI-1000 Ljubljana, Slovenia; 3D.I.I.V. Ltd., for marine, freshwater and subterranean ecology, Obala Petra Lorinija bb, HR-23281 Sali, Croatia * corresponding author ([email protected])

The endemic Mediterranean seagrass Posidonia oceanica (L.) Delile is a very important benthic primary producer. Even though its meadows have multiple ecological functions they have been modified and destroyed due to a changing environment and an increased human impact. As a slow-growing and long-living species sensitive to environmental changes, P. oceanica is a good bioindicator. In this work, phenolic compounds (total phenolics, flavonoids and anthocyanins), photosynthetic pigments and leaf morphology were evaluated in intermediate leaves of P. oceanica. The samples were collected on 2 (or 3) and 10 m of depth, in summer and winter on three locations of the Zadar coastal area: Zadarski kanal (ZK), Brbišćica Bay (B) and Zaglav Port (ZA) on island Dugi otok. The aim was to find correlation among measured parameters, environmental conditions and the primary production of the seagrass meadow and to estimate the suitability of parameters to be used as bioindicators. Plants collected in ZK and B at 10 m depth had higher portion of flavonoids in total phenols than those taken at 2 (3) m. Also, higher values were noticed in winter. In samples from ZA the portion of flavonoids was almost the same regardless of depth and season. On the other hand, anthocyanin portion was the highest in summer at 2 (3) m of depth. The ratio of chlorophyll a and chlorophyll b was higher in winter in plants grown at both depths and all locations. The ratio of carotenoids and total chlorophylls was mainly 0.20-0.23, except in ZK in summer at 2 m with significantly lower value (0.16). Similar results were obtained for ZA samples. In morphological study mesophyll width was between 74% and 88% of total leaf thickness. Plants at all three locations had thicker mesophyll in winter and at 2 (3) m depth but the differences were not always significant. Principal component analysis showed that samples grouped mostly due to season and depth. Two deviations were noticed, namely samples in ZA at 3 m and in winter were closer to ZA samples collected at 10 m. Also, samples from ZK at 2 m and in summer differed from other samples due to higher primary production which could be connected with human impact.

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Temperature affects reproductive traits in olive tree (Olea europaea L.) – flowering timing and pollen performance

Gabriela Vuletin Selak1*, Slavko Perica1, Smiljana Goreta Ban2 1 Institute for Adriatic Crops, Put Duilova 11, 21 000 Split, Croatia 2 Institute of Agriculture and Tourism Poreč, K. Hugues 8, 52 440 Poreč, Croatia * corresponding author ([email protected])

The reproductive phase has proven to be one of the most temperature sensitive stages in plants. Relatively high or low temperatures are often correlated with low fertilization percentages and low fruit setting. The interest in the effect of temperature on reproductive processes is increasing because the global rise in temperature has affected plant populations by inducing a shift in several phenological traits, such as dormancy release and flowering time. In the Mediterranean basin, olive flowers between April and June which is later than most other tree species. This later date allows olive to meet the requirements of an ample accumulation of heat units during the growing season. Six olive cultivars (‘Lastovka’, ‘Leccino’, ‘Levantinka’, ‘Oblica’, ‘Pendolino’ and ‘Itrana’) were employed in field experiment during three years to determine the effect of accumulated temperatures preceding the flowering periods on the beginning of flowering. The flowering periods were assessed by visual examination of eight trees per cultivar. The average flowering date (AFD) was calculated as the number of days from 1 January until the start of the flowering period. The AFD was determined for each experimental year using flowering dates of all tested cultivars. The number of chilling units (CU) was calculated using the daily maximal and minimal air temperatures. The heat accumulation start dates were the first days after the chilling requirement had been met and continued until the start of flowering season. Heat units were expressed in growing degree-days (GDD). The effect of temperature on pollen germination and pollen tube growth was evaluated using in vitro system. Pollen performance has been tested under different temperatures (15, 22±2, 30, 35 and 40 ºC) to evaluate the sensitivity of the microgametophyte to temperature extremes. The olive tree can be used as a sensitive biological indicator of climatic changes since the yearly temperature variations control the timing of the flowering period. High correlations have been found between values of AFD and those of CU and GDD. Moreover, simultaneous flowering of different cultivars is important in achieving a high fruit set after cross-pollination. The flowering periods of most of the studied cultivars overlapped to some degree. Pollen germination in vitro was enhanced when pollen grains were exposed to higher temperatures up to 35 ºC; however germination failed at 40 ºC. In most of the studied cultivars, the longest pollen tubes were developed at 22±2 ºC and the pollen tube length decreased with higher incubation temperatures. Temperature significantly affected flowering timing and pollen performance in olive cultivars and therefore participates in regulation of final fruit set and yield.

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Examination of physiological response of common bean (Phaseolus vulgaris L.) to drought stress

Mateja Zupin1*, Marko Maras1, Jelka Šuštar Vozlič1, Marjetka Kidrič2, Dominik Vodnik3, Jaka Razinger4, Vladimir Meglič1 1Agricultural Institute of Slovenia, Crop Science Department, Slovenia 2Jožef Stefan Institute, Department of Biochemistry and Molecular Biology, Slovenia 3University of Ljubljana, Biotechnical faculty, Department of Agronomy, Slovenia 4Agricultural Institute of Slovenia, Plant Protection Department, Slovenia *corresponding autor ([email protected])

Common bean is nutritionally very important legume plant that exhibits sensitivity to drought which affects its growth and yield. Plants developed mechanisms in adapting to drought, which are expressed by changes of gene expression and functional protein content, together with responses at physiological and morphological levels. Identification of changes in response to drought in different genotypes will provide markers essential for introduction and implementation of marker assisted selection in breeding for greater drought tolerance. To establish differences between plants subjected to water withdrawal and normally watered plants of common bean, the parental cultivars, drought tolerant Tiber and drought susceptible Starozagorski čern, were grown in a growth chamber under controlled environmental conditions. Plants were watered daily to the same pot weight. After three weeks the half of plants were stressed by withholding irrigation. Physiological parameters were measured on third trifoliate leaves at different drought stages. The hydration state of leaves was defined by their relative water content (RWC), water potential (Ψw) and different photosynthetic parameters. Detached leaf samples are kept at -80°C for further analyses. In water stressed plants RWC dropped to 50% compared to control plants. Due to the reduction of water potential, the duration of the leaf stomata closure was increased as confirmed by the results of photosynthetic and other physiological parameters. Other differences in response to drought between the cultivars were identified, which is of great importance for further study of genetic factors governing drought response by means of QTL mapping.

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Analyses of salt stress alleviation with homobrassinosteroid in germinating barley roots

Sevgi Marakli*, Nermin Gozukirmizi Istanbul University, Faculty of Science, Department of Molecular Biology and Genetics, 34134, Vezneciler, Istanbul-TURKEY * corresponding author ([email protected])

Effects of Homobrassinolide (HBR) (0.5-1 μM HBR) on root germination and cell division of barley (Hordeum vulgare L. cv. Hilal) roots grown under different salt concentrations (150 mM and 250 mM) for 48 and 72 h were investigated with their controls. Increased salt concentration caused negative effect on primary root and seminal root lengths as compared to control (1.8 ± 0.6 and 2.4 ± 0.4, 48 h and 72 h, respectively), the lengths of primary root decreased in 150 mM salt concentration (1 ± 0.4 and 1.5 ± 0.6, 48 h and 72 h, respectively) and 250 mM (0.5 ± 0.2 and 0.9 ± 0.2, 48 h and 72 h, respectively) salt concentration. On the other hand, HBR (0.5-1 μM HBR) applications enhanced primary root lengths, seminal root lengths and number of roots from one seed compared to controls and only salt-treated samples. Salt + HBR treatments showed varying results depending on concentration and timing. Mitotic activity in control barley roots was 20.4% and 23.6% (48 h and 72 h respectively) of the cells. Salt stress reduced mitotic index (8.3% and 5.6% in 150 mM salt treated and 5.5% and 3.6% in 250 mM salt treated samples for 48 h and 72 h). Moreover, mitotic index of HBR application was higher than both control and salt treated samples. Increasing concentration of HBR alleviated salt stress effectively but their effect could be seen after long time (1 μM application showed more mitotic index for 150 mM salt and 250 mM salt application both at 48 h and 72 h). Roots treated with HBR showed more mitotic activity, mitotic abnormalities and significant enlargements at the root tips. In addition, salt + HBR applications stimulated root lengths and showed more mitotic activity and mitotic abnormalities when compared to only salt-treated samples. Depending on concentration and timing, HBR hormone alleviates salt stress on germinating barley roots.

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Polytrichaceous mosses are resistant to short termed salt stress

Marija Rajčić, Vesna Nerić, Milorad Vujičić, Marko Sabovljević, Aneta Sabovljević*

Institute of Botany and Garden, Faculty of Biology, University of Belgrade, Takovska 43, 11000 Belgrade, Serbia * corresponding author ([email protected])

Bryophytes are among the earliest land plants. They have a high degree of abiotic stress tolerance, which helps them to survive in the harsh environment. The aim of this study was to investigate the effects of different salt concentrations (5mM - 500mM NaCl) on biomass and photosynthetic pigment (chlorophylls and carotenoids) content in selected bryophyte species (Atrichum undulatum (German and Serbian genotype) and Polytrichum formosum (German genotype). These species were cultured for 4 days in vitro, at the half strength Murashige and Skoog (MS) medium enriched with different NaCl concentrations and subsequently 24 days at the basal MS half strength medium. At higher concentrations of NaCl, total chlorophyll content is decreasing. Index of multiplication at lower NaCl concentrations is linearly decreasing. Survival of plants (all three genotypes) is 100% up to 100mM NaCl. At higher concentration of NaCl (200-500 mM) there is linearly decrease of the survival rate. Chlorophyll content increased up to 200 mM NaCl and carotenoid content up to 300 mM NaCl, both in comparison to the untreated control group. Biomass production in both species, decreased in the presence of higher concentrations of salt, compared to biomass production of control groups. Although unexpected, both species and three genotypes showed resistance to salt stress.

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Rain simulation with heavy metal deposition: effects of exposure duration and lead concentration on survival and development of the moss Hypnum cupressiforme

Marko Sabovljević1*, Mitja Skudnik2, Milorad Vujičić1, Jovana Pantović1, Nada Nikolić1, Zvonka Jeran3, Franc Batič4, Aneta Sabovljević1 1Institute of Botany and Botanical Garden, Faculty of Biology, University of Belgrade, 11000 Belgrade, Serbia 2Slovenian Forestry Institute, Department of Forest and Landscape Planning and Monitoring, Večna pot 2, 1000 Ljubljana, Slovenia 3”Jožef Stefan” Institute, Department of Environmental Sciences, Jamova 39, 1000 Ljubljana, Slovenia 4Biotechnical Faculty, Universiy of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia * corresponding author ([email protected])

The moss Hypnum cupressiforme Hedw. is widely used as a model-moss in study of atmospheric deposition of contaminants. Many European-wide heavy metal deposition measurements in mosses as biomonitors were done on this ubiquitous moss. However, up to date there was no experimentation on this moss in controlled axenical conditions. With the aim to control the effects of rain duration and lead concentration in simulated rain on mosses we established in vitro culture of moss Hypnum cupressiforme. This pleurocarpous moss was grown in controlled light and temperature conditions. Axenically propagated plants over a few years in laboratory conditions were used as start material for further experimentation. Forty explants (each of 10 mm long) per each exposure period and concentration were exposed to simulated rain for 0.5, 2 and 24 hours, and the rain contained 0, 50, 150, 300 μM Pb-acetate as a carrier of lead. After exposure to simulated rain the plants were grown for eight weeks on half strength Murashige and Skoog basal solid medium prior to developmental features and survival measurements. The results obtained clearly showed that the duration of exposure to the simulated rain and the increase of Pb concentration decreased the regeneration of the 10 mm start explants both in secondary protonema production and index of multiplication. Survival also decreased with duration of exposure and increase of Pb concentration. The lethal concentrations were those above 700 μM (tested sequently up to 5mM) of Pb-acetate in any duration of simulated rain tested. Further experiments of simulated rain to chlorophyll production and other pigment contents, as well as synergism with other heavy metals and the repetition of subsequent simulated rains followed by the intermediate time of moss recovery are ongoing.

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The effect of nickel stress on snake's head fritillary (Fritillaria meleagris L.)

Valerija Vujčić1*, Marija Petrić2, Sandra Radić Brkanac1, Slađana Jevremović2, Angelina Subotić2, Branka Pevalek-Kozlina1

1 Department of Biology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, Zagreb, Croatia 2 Institute for Biological Research "Siniša Stanković", University of Belgrade, Bulevar despota Stefana 142, Belgrade, Serbia * corresponding author (vvujcic©biol.pmf.hr)

Snake's head fritillary (Fritillaria meleagris L.) is an endangered and protected plant species in the family Liliaceae mainly inhabiting moist grasslands. As a result of negative anthropogenic impacts such as ameliorization and urbanization, the habitats rapidly disappear and/or are affected by pollutants such as heavy metals. The aim of the study is to estimate level of sensitivity and/or resistance of snake's head fritillary to nickel (Ni), a metabolically important microelement though also toxic when present at excessive amounts in soil. Plants cultivated in vitro were subjected to increasing Ni concentrations (0-5 mM) for four weeks. Relative growth rate (RGR) and certain indicators of oxidative stress - soluble proteins, malondialdehyde content and antioxidant enzyme activities were measured. A decrease of RGR based on fresh weight and protein content was noted in response to growing Ni concentrations whereas malondialdehyde (indicator of lipid peroxidation) content increased. Antioxidant response was observed under Ni stress, with large increases in catalase and peroxidases. The results suggest induction of oxidative stress under Ni toxicity, but also demonstrate capacity of F. meleagris to upregulate its antioxidative defense, thus indicating certain degree of plant's tolerance to Ni exposure.

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Analysis of stem proteome of common bean under drought using in-gel stable isotope labeling

Tanja Zadražnik1*, Jelka Šuštar-Vozlič1, Wolfgang Egge-Jacobsen2

1 Agricultural Institute of Slovenia, 1000 Ljubljana, Slovenia 2 University of Oslo, Department of Molecular Biosciences, Oslo, Norway * corresponding author ([email protected])

Drought is an abiotic stress that strongly influences plant growth, development and productivity. By proteomics study it is possible to identify the complex mechanism of water-stress response. The aim of this research was the analysis of common bean (Phaseolus vulgaris L.) stem proteome changes under stress conditions by applying a water deficit treatment. Quantitative proteomics using in-gel stable isotope labeling of stem proteins in combination with mass spectrometry was used for the analysis. Mass spectrometry data were analyzed using MaxQuant software, where quantification and presentation of the results were performed. In total, 45 protein groups were found to change significantly in abundance between control and stressed samples. The quantified proteins were grouped into several functional groups, mainly into energy metabolism, photosynthesis, proteolysis, synthesis and proteins related to ROS, defence and stress. Mixed abundance profiles of proteins involved in energy metabolism and photosynthesis showed different range of proteins involved in the complex regulation and production of different metabolites within energy metabolism and reaction of photosynthesis. Heat shock protein showed the highest increase in abundance among all 45 proteins, suggesting its role in protecting plants against stress by re-establishing normal protein conformation and thus cellular homeostasis. Proteins involved in protein synthesis were all increased in abundance under drought. Increased levels of protein synthesis are important in order to restore the damaged proteins for full restoration of the plant cell's metabolic activities and general growth. A decrease in abundance of protein tubulin, a classical cytoskeleton-related protein was noticed. This indicate the involvement of cell size adjustment as one of the cellular adaptive strategies to drought stress. Also other important proteins were quantified and were included in a wide range of biological pathways that are involved in plant protection. The research provided the basic insight into the molecular regulatory mechanism of stem proteins in common bean under drought.

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3 Moderne tehnologije in Rastlinska biotehnologija

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Green mathematics: Benefits of including biological variation in your data analysis.

L.M.M. Tijskens 1,*, Rob E. Schouten 1, Tatjana Unuk 2, Marjan Simčič 3 1 Wageningen University, Horticulture and Product Physiology, The Netherlands 2 University Maribor, Faculty of Agriculture and Life Sciences, Slovenia 3 University of Ljubljana, Biotechnical Faculty, Slovenia * corresponding author ([email protected])

Variation exists in all samples of natural products. Sometimes the variation is small, often it is large. To

avoid interference from this biological variation, the main effort in developing handling procedures in

horticulture, is to try to make variation in production during growth as small as possible while

maximising the yield. The still existing variation is dealt with in various postharvest sorting and grading

systems as used in packing houses. That works most of the time pretty well for commercial purposes. So,

this technology is applied for the last 100 to 150 years quite successfully.

This strategy, however, does intrinsically not invite to study the basic rules that govern this biological

variation and prevents thereby a more basic understanding of its sources and dynamic behaviour in time.

It therefore also prevents efforts of turning that omnipresent nuisance into an advantage. Understanding

the dynamics of biological variation (also in QTLs and other genetic information), will open complete new

alleys for optimising production and distribution of horticultural products. Knowledge on variation in

product properties, well in advance of harvest time, enables a more suitable use of product classes in the

postharvest supply chain.

In this paper, the methodology, statistical as well as mathematical, of including differences between

individuals in a batch of products will be explained (Tijskens et al. 2005) and highlighted (Tijskens et al

2003), and the benefits in terms of reliability of data interpretation will be indicated based on a number

of examples, ranging from colour and size development in the apple orchard in Slovenia.

Tijskens L.M.M. Konopacki P., Simčič M. (2003). Postharvest Biology & Technology 27/1, 15-25. Tijskens L.M.M., Heuvelink E., Schouten R.E., Lana M.M., van Kooten O. (2005). Acta Horticulturae 687, 39-46 Groups. Fourth Edition, Springer, Berlin: 506 pp.

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Infrared spectroscopy in plant science

Meta Kokalj*, Samo Kreft University of Ljubljana, Faculty of pharmacy, Aškerčeva 7, 1000 Ljubljana, Slovenia * corresponding author ([email protected])

Infrared spectroscopy (IR) has been applied in plant material identification, in assessing quantity of certain compounds and also imaging. IR is fast, easy to use and does not use organic solvents. In order to apply IR to plant samples it is necessary to combine it with chemometrical methods. Infrared spectra are a very large and complex sets of data therefore the analysis is very complex. However once the optimal model for given data is established, the use is very simple and can be automatized (Bunaciu A. A. 2011, Gad H. A. 2011). The analysis with IR is composed of three main steps: spectra measurement, spectra pre-treatment and chemometrical analysis. Each of these steps has to be optimized case-bycase since the properties of the samples and the goal of analysis can be very different. In different modes of spectra measurement it is important to take in account from which part of the sample (the surface or the inside) the spectra are collected, (Kokalj M., Kolar J., et al. 2011). With the pretreatments of the spectra important information can be emphasized or, if inappropriate technique is used, important information can be lost (Kokalj M., Rihtarič M., et al. 2011). Since the infrared spectra are a complex set of data it is also important to carefully choose the mathematical and statistical methods for their interpretation (Kokalj M.,2010). There are no general rules, so optimal conditions have to be found for each specific case. Bunaciu A: A., Aboul-Enenin H. Y., Fleschin S. (2011). Applied Spectroscopy Reviews 46, 251-260. Gad H. A., El-Ahmady S. H., Abou-Shoer M., Al-Azizi M. M. (2011). Phytochemical analysis 24, 1-24. Kokalj M., Kolar J., Trafela T., Kreft S. (2011). Phytochemical analysis

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